Pre-equilibrate dishes: Incubate the dishes with culture medium. Pipet 2-3 ml of medium into the 35 mm dishes or 3-4 ml into the 50 mm dishes and incubate at 37° C for 15 minutes.

Add cell suspension to microwell: Remove the culture medium by aspiration and plate cells onto the glass surface. Pipet 250 µl of the cell suspension (cells suspended in culture medium) into the 10 mm diameter microwells, 500 µl of cell suspension into the 14-mm microwells, or 1 ml of cell suspension into the 20-mm wells. Incubate the dishes for 1 hour at 37° C.

Add additional medium: After 1 hour, gently fill the remainder of the dish with medium. Add 2-3 ml to the 35 mm dishes or 3-4 ml for the 50 mm dishes.

Note: After the initial one hour period to allow cells to attach to the glass surface, it is important to fill the dish to normal levels in order to minimize the effects of evaporation and to avoid inducing changes in osmolarity.

Q2: What type of glass bottom dish should I use to grow my cells?

It is hard to predict which type of glass bottom dishes (uncoated, poly-d-lysine coated, or collagen coated) will work best with your specific cell type. Many transformed or cancerous cell lines will grow on uncoated dishes. Poly-lysine coated dishes work well for neuronal culture and for many primary cells; other cells prefer a collagen coating. Also, many researchers purchase our uncoated dishes and apply their own specialized coating. You can also Google search for published methods utilizing glass bottom dishes and your cells.

Note: For almost all microscopy applications, the No. 1.5 coverslip thickness is preferred.

Q3: How can I search for published methods utilizing MatTek's glass bottom dishes?

We have collected and catalogued hundreds of technical papers that cite the use of MatTek Glass Bottom Culture Dishes. You can search our database of cataloged publications on our website. Note: If the exact glass bottom dish part number is unspecified in a literature article, it is safe to assume that uncoated dishes were used (e.g. Part #’s: P35G-1.5-14-C, P50G-1.5-30-F, etc.)

Q4: What should I do if my cells won't grow or if I want to improve growth in the glass bottom dishes?

Although our poly-d-lysine or collagen coating works well for the vast majority of cells, some customers find it necessary to coat the glass bottom dishes themselves. They purchase the uncoated dishes and use the following hydrochloric acid (HCl) pretreatment along with their coating of choice.

After 15 minutes, decant the HCl and rinse the dish 3x with phosphate buffered saline (PBS) and 2x with ultrapure H2O.

Apply the coating to the dishes.

Add a similar volume of the medium in which you will plate your cells to pre-equilibrate the glass surface. Incubate the medium in glass bottom dishes for 15 min at 37°C. Remove the medium and then plate your cells.

Yes, but for most applications, cells growing in the glass bottom dish can be viewed without removal of the coverslip using a variety of inverted and upright microscopes.

If necessary (e.g. for long-term storage purposes), the coverslip can be removed using the following procedure:

Order Part # PDCF OS 30 (Fluid for removal of coverslips from glass bottom dishes)

Invert the cover of the dish.

Pipette 1.0 ml of fluid into the inverted cover.

Place the bottom of the dish onto the cover. Make sure that the liquid in the inverted cover is touching the bottom of the coverslip.

Allow the dish to sit in the fluid for 45 minutes at room temperature.

Dry the bottom of the coverslip with an absorbent paper towel.

Place the dish on a clean surface. Using forceps, press down on the inner edge of the coverslip to separate the coverslip from the dish.

Note: If the above procedure is followed, the PDCF OS 30 fluid will not contact the cells and will not disrupt cells on the coverslip or the staining thereof. Coverslips can be removed without breakage.

Q6: How can I control the temperature of the glass bottom dishes for in situ (live) microscopy?

In order to approximate physiological conditions, the temperature of the medium contained within the glass bottom dishes can be controlled by using a microscope stage heater and an appropriate stage adapter.

Q7:Why would one want to use glass bottom dishes containing gridded coverslips? What is the size of the grid? What if I can't see the grid?

The gridded coverslips allow one to refer to specific cells and follow them over time. For instance, individual cells can be microinjected, returned to the incubator, and observed at multiple time points since each cell can be identified with a unique alpha-numeric coordinate in the dish. Glass bottom dishes containing gridded Bellco Glass coverslips are available. Standard gridded dishes are designated as Part #’s: P35G-1.5-14-CGRD and P50G-1.5-14-FGRD.

Grid size: The grid on Part #’s P35G-1.5-14-CGRD and P50G-1.5-14-FGRD consists of 520 unique alphanumeric squares. Each square measures 600 microns x 600 microns. The line thickness is 20 microns.Visualization of the grid: The grid should be readily observable using a 10X brightfield objective. After you locate the cell of interest, you can switch to a higher magnification or fluorescence objective. However, the grid will not be observable using higher power or fluorescence objectives.I can’t see the grid: A confluent monolayer of cells will typically mask the grid making it difficult or impossible to visualize. If your cells are confluent, you can utilize Part # P35G-1.5-14-CGRD-D which places the grid on the outside of the dish where it is unaffected by the cells growing on the coverslip. To use this dish, find a cell of interest and then focus down to the bottom side of the coverslip to get its coordinates.

Q8:How can I use the glass bottom dishes to look at tissue slices?

The glass bottom dishes can also be used to image tissue slices. The slices are adhered to the dishes using Corning Cell-Tak. A research paper utilizing MatTek’s glass bottom dishes to perform confocal microscopy on brain slices is available.

Q9: Can the glass bottom dishes be re-used?

We do NOT recommend re-using the glass bottom dishes. The surface properties of the substrate on which cells are cultured have a profound effect on cell structure and function. Re-use of dishes will introduce uncontrolled variables into your experiments which may affect the phenomenon under study.
MatTek glass bottom dishes are meant for single-use experiments.

Microscopy Types and Techniques

Q10: Are the glass bottom dishes good for confocal and fluorescence microscopy?

Yes. The glass bottom dishes are excellent for both confocal and fluorescent microscopy. Important glass properties are:

Incident ultraviolet rays with wavelengths longer than 320 nm do not cause fluorescence.

Mercury lines at 334 and 365 nm do not create auto-fluorescence. (Note: For mercury illumination, filter out the mercury lines with wavelengths shorter than 313 nm to obtain best possible results.)

Refractive index (@ 20°C): nd= 1.5230 tolerance ± 0.0015

Abbe number V = 55.

Q11: How can I use the glass bottom dishes for Nomarski Differential Interference Contrast (DIC) microscopy?

You will need to order BOTH a glass bottom dish and a glass cover. Order any P35G dish (e.g. Part #’s: P35G-1.5-14-C or P35G-1.5-20-C) along with a glass cover (Part #: P35GTOP-0-20-C). Both the glass bottom and glass cover are necessary so that the entire light path travels only through high optical quality glass. The glass covers can be re-used following resterilization by soaking them in 70% ethanol for 30 minutes. The covers CANNOT be autoclaved.

Q12: Can the dishes be used for high-resolution techniques such as GSDIM, dSTORM, PALM , or TIRF microscopy?

Yes. For high-resolution techniques such as GSDIM, dSTORM, PALM, TIRF, and for any objective with a numerical aperture > 0.7, tight control of the coverslip thickness is crucial. Therefore, use of Part # P35G-0.170-14-C dish which utilizes a high tolerance No. 1.5 coverslip (0.170 ± 0.005 mm) is necessary.

Q13: Why are the 50 mm glass bottom dishes useful for microinjection and maintaining a constant atmosphere in the dish?

The 50 mm glass bottom dishes (part #’s beginning with P50G-) are useful for:Microinjection: The larger diameter (50 mm) and the lower side wall (9 mm) allows easier access to cells in microinjection experiments.Atmosphere maintenance: The 50 mm dish has a cover that snaps onto the dish bottom and thereby prevents loss of the 5% CO2 atmosphere while the dish is out of the incubator. This can be important for experiments in which dishes will be observed for extended periods.

Q15: What coverslip thickness should I use? What do the various coverslip numbers correspond to?

Almost all objectives are optimized for a No. 1.5 coverslip thickness. Use of the No. 1.5 thickness gets increasingly important for higher numerical aperture coverslips (NA > 0.7). Use of other coverslip thicknesses can lead to optical distortion and loss of resolution. Therefore, for the vast majority of microscopy applications, the No. 1.5 coverslip thickness is optimal (e.g. Glass bottom dish Part #s: P35G-1.5-xx-C, P50G-1.5-xx-F, P60G-1.5-xx-F, etc.).

Q16: What is the adhesive used to attach the coverslips to the petri dishes?

Although the specific identity of the adhesive is proprietary, the adhesive used is a non-toxic silicone that has been shown to be compatible with a broad variety of cells including primary neurons and many other difficult-to culture, fastidious cells.

Q17: How long can the dishes be stored?

Coated glass bottom dishes can be stored in the dark at room temperature for up to 1 year. Uncoated dishes can be stored for up to 5 years without any decline in cell growth properties.

Q18: Over what temperature range can the glass bottom dishes be used?

The glass bottom dishes can be used over the temperature range -20°C to +45°C. The dishes will become disfigured at 55°C (131°F) and above. The glass bottom dishes CANNOT be autoclaved.

Q19: What are properties of the glass used in the glass bottom dishes?

MatTek uses the highest quality, borosilicate German glass coverslips in its glass bottom dishes. The coverslip properties are as follows:

Highest hydrolytic resistance (hydrolytic class 1).

Excellent resistance to chemicals.

Emission of alkali approximately 15 to 24 µg Na2O/g glass.

Excellent properties for fluorescent microscopy.

Incident ultraviolet rays with wavelengths longer than 320 nm do not cause fluorescence.

Mercury lines at 334 and 365 nm do not create auto-fluorescence.(Note: For mercury illumination, filter out the mercury lines with wavelengths shorter than 313 nm
to obtain best possible results.)

Refractive index (@ 20°C): nD ** = 1.5230 tolerance ± 0.0015.

Abbe number V = 55.

Q20: How deep is the micro-well of the glass bottom dish?

The depth of the micro-wells in the glass bottom dishes depends on the type of dish and the thickness of the dish bottom as follows:

Corning 35 mm: 0.70-0.75 mm

Falcon 50 mm: 1.00-1.10 mm

Falcon 60 mm: 1.15-1.20 mm

Falcon 100 mm: 0.90-1.00 mm*

Falcon 6-well plate: 1.45-1.55 mm

Falcon 12-well plate: 1.45-1.55 mm

Falcon 24-well plate: 1.10-1.20 mm

Falcon 96-well plate: 1.05-1.25 mm

Q21: What is the chemical compatibility of Glass Bottom Dishes and Multi-Well Plates? Can they be used with organic solvents?

The body of the glass bottom dishes and multi-well plates is made from polystyrene. Therefore, they have limited compatibility with organic solvents. Please see the chemical compatibility table.

Solvent

Chemical Compatibility

Acetone

Poor

Ammonium hydroxide (1N)

Fair

Ammonium hydroxide (25%)

Fair

Aniline

Good

Butanol

Good

Chloroform

Poor

Dimethylformamide

Poor

Dimethylsulfoxide(DMSO)

Poor

DMSO/H2O (20/80)

Good

Dioxane

Poor

Ethanol

Good

Hexane

Poor

Hydrochloric acid (25%)

Good

Hydrochloric acid (concentrated)

Fair

Methanol

Good

Methyl ethyl diketone

Poor

Methylene chloride

Poor

Nitric acid (25%)

Poor

Nitric acid (concentrated)

Poor

Sodium hydroxide

Good

Toluene

Poor

Xylene

Poor

Q22: How do I know that the glass bottom dishes are sterile?

All glass bottom dishes are gamma irradiated at an FDA approved and certified vendor. We sterilize our dishes in bulk and typically >2000 separate cases are sterilized at the same time. Since sterility is an absolute requirement for all of our customers, the gamma dose that we use is excessive in order to ensure sterility. Following sterilization, dishes are subjected to quality control analysis to verify sterility: they are incubated in antibiotic- and anti-fungal-free medium for 7 days. In addition, each box has a gamma irradiation indicator that turns red upon exposure to gamma rays.

Q23: How do the high tolerance P35G-0.170-14-C dishes differ from the standard P35G-1.5-14-C? For what applications would the P35G-0.170-14-C dishes be useful?

The P35G-0.170-14-C dishes will improve picture quality (see Figure below) versus the P35G-1.5-14-C dishes for any high numerical aperture objective used in confocal, fluorescence, GSDIM, dSTORM, PALM total internal reflection (TIRF), and other high numerical aperture objectives. For example, quantitative measurements using P35G-0.170-14-C dishes gave z-resolution of +/- 9.5% while z-resolution in the P35G-1.5-14-C dishes gave z-resolution of +/- 17.3% (n=5).

A

B

Figure: Improved Z-axis resolution – Effect of high tolerance glass coverslips (in P35G-0.170-14-C glass bottom dishes) on imaging of sub-resolution beads using:A) P35G-0.170-14-C and B) P35G-1.5-14-C glass bottom dishes.
5 µl of FITC labeled 175nm PS-Speck sub-resolution beads were added to well of the glass bottom dishes and allowed to dry. After drying, 200 ul of water were added and the beads were imaged using a Zeiss LSM510 confocal microscope equipped with an Olympus UPLSAPO 60x (NA=1.2) water immersion objective.

Q24: What is the molecular weight range of the poly-d-lysine used to coat the PDL coated glass bottom dishes?

The poly-d-lysine used to coat the P35GC-x-xx-C and P50GC-x-xx-F glass bottom dishes is in the molecular weight range of 70,000-150,000 Daltons.

Q25: Why are the dishes coated with poly-d-lysine (instead of poly-l-lysine)?

Both untreated glass and cells are negatively charged. Poly-lysine is applied to the glass surface to make it positively charged, thereby increasing electrostatic attraction between the glass surface and the cells and thus improving cell attachment. Poly-d-lysine is favored because the d-enantiomer is less prone to protease-mediated breakdown than the naturally-occurring l-enantiomer. Otherwise, Poly-d-Lysine and Poly-l-Lysine are equivalent.

Q26: What type of collagen is applied to the collagen coated glass bottom dishes (part #: P35GCol-x-xx-C or P50GCol-x-xx-F)?

The collagen used to coat the P35GCol-x-xx-C or P50GCol-x-xx-F glass bottom dishes is type 1 rat tail collagen.

Q27: Do the poly-lysine or collagen coatings affect the optical properties of the glass bottom dishes?

The coatings are monolayer coatings which do not affect the optical properties of the glass bottom dishes.

Special Orders

Q28: Other than the standard 10 mm, 14 mm, and 20 mm diameter glass surfaces, do the glass bottom dishes come with any alternative diameters?

Glass bottom dishes with 7-mm diameter glass surfaces are standard products (e.g. Part # P35G-1.5-7-C or P35G-0-7-C). In addition, 50-mm, 60-mm- and 100-mm dishes come with a 30-mm diameter glass surface (e.g. Part #P50G-1.5-30-F, P60G-1.50-30-F, P100G-1.5-30-F) and are standard products; these dishes maximize the surface area for cell growth. When very expensive reagents need to be conserved, dishes with a 5-mm diameter glass surface (e.g. Part #: P35G-1.5-5-C) are also available on a special order basis.

Q29: What is the lead-time necessary for special orders? Is there a special order charge? Can special order items be returned?

Lead-time: Special order items typically can be produced within 2-3 weeks of receipt of a purchase order. However, bulk sterilization of the glass bottom dishes occurs only once every 6 weeks. Therefore, the lead-time for sterilized, special order products can vary between 3-8 weeks.Note: Lead-times can be shortened if the customer will sterilizes the dishes using UV and/or 70% ethanol (e.g. 40 mins under UV light in a tissue culture hood and/or immersion if 70% ethanol for 30 minutes.Special order charges: For special orders of ≥ 3 cases, there is no special order charge. However, for special orders of < 3 cases, a special order charge is assessed per case.Return policy: Special orders items cannot be returned.

Q30: What are the advantages of using multi-well glass bottom plates compared to the standard glass bottom culture dishes?

A: The main advantage of the multi-well glass bottom plates is that you can grow 6, 12, 24, 48 or 96 cultures under identical conditions in the same culture plate. The multi-well plates are ideal for high throughput, high/high content screening applications.
B: Analysis using the multi-well plates is streamlined because only one (1) plate (versus multiple petri dishes) needs to be handled.
C: For a number of applications, treatment of the cultures (e.g., irradiation) is simplified using the multi-well plates.
D: Smaller wells in the glass bottom multi-well plates are useful for application of precious reagents in smaller volumes.

Q31: Do you offer multi-well glass bottom plates that are black?

Yes. In order to minimize back-scattered light and background fluorescence, we offer 96-well glass bottom plates (part # PBK96G-1.5-5-F) that are black. All properties of the PBK96G-1.5-5-F plates are identical to the standard, clear-wall 96-well glass bottoms plates (Part #: P96G-1.5-5.F) except the sidewalls of each well in the PBK96G-1.5-5-F plates are opaque (black). Note: Back-scattered light and background fluorescence is not a problem in the larger well 6-well, 12-well, 24-well, and 48-well glass bottom plates.

3D Tissue Models

Q1: How long has MatTek been in business?

MatTek Corporation was founded in 1985 by two MIT chemical engineering professors. In 1991, the company leveraged its core polymer surface modification technology into the emerging tissue engineering market. In 1993, EpiDerm, MatTek’s first in vitro human tissue equivalent, was introduced and has been in continuous production from 1993 to the present.

Q2: What are MatTek's major product lines?

MatTek produces a variety of normal (non-transformed), human cell-derived, 3-dimensional, organotypic in vitro tissue equivalents. This Web site, MatTek.com, is devoted to providing detailed information about our in vitro human tissue equivalents. They are mitotically and metabolically active, closely mimic their in vivo counterparts, both structurally and biochemically, and do so with guaranteed reproducibility.

MatTek also produces a line of Glass Bottom Culture Dishes. These dishes are used in confocal, polarized light, and fluorescence microscopy techniques, and are ideal for live cell microscopy applications.

Q3: How many different types of in vitro human tissue equivalents does MatTek produce?

MatTek produces 9 in vitro human tissue equivalents, all of which are derived from human epithelial cells (click on the tissue name for detailed information):

EpiDerm, our human epidermal tissue equivalent, consists of normal, human cell-derived epidermal keratinocytes that have been cultured to form a multilayered, highly differentiated model of the human epidermis. EpiDerm, also known generically as Reconstructed Human EpiDermis (RhE), has been in continuous production for over 15 years. There is also an “under-developed” version, EpiDerm-201

MelanoDerm is a human skin equivalent composed of keratinocytes and melanocytes that have been cultured to form a multilayered, highly differentiated model of the human epidermis.

Melanoma is a full thickness melanoma skin model composed of human malignant melanoma cells, normal, human-derived epidermal keratinocytes and normal, human-derived dermal fibroblasts that have been cultured to form a multilayered, highly differentiated epidermis with melanoma cells at various stages of CM malignancy.

EpiOcular, our corneal model, consists of normal, human cell-derived epidermal keratinocytes that have been cultured to form a stratified, squamous epithelium similar to that found in the cornea.

EpiAirway consists of normal, human cell-derived tracheal/bronchial epithelial cells that have been cultured to form a pseudo-stratified, highly differentiated model that closely resembles the epithelial tissue of the respiratory tract.

Tissues are also available in 24-well and 96-well high thru-put plates. Special configurations are also available for specific tissue types (ex. EpiAirway tissues produced in “snapwell” culture inserts for use in Ussing Diffusion Chambers).

Q6: Do any of the in vitro human tissue equivalents also contain Dendritic/Langerhans Cells?

Yes. EpiVaginal with Langerhans Cells (“VLC” products) is a released product. Several other MatTek in vitro human tissue equivalents that incorporate Dendritic Cells are in “beta” phase. Please contact MatTek for more detailed information.

Q7: What are some typical applications for MatTek's in vitro human tissue equivalents?

An example of a pharm/tox application of MatTek’s in vitro human tissue equivalents is as follows: after a new drug library has been processed thru a biochemical-based (enzyme) primary screening and a cell-based secondary screening, but PRIOR to performing pre-clinical animal studies, the remaining drug candidates are passed through a tertiary in vitro human tissue-based screening to determine optimum permeability and/or minimal toxicity characteristics. Those drug candidates that pass this tertiary screening then move onto the animal-based study, but that study will now require fewer animals, and can therefore be structured as a small confirmatory study to meet FDA (or other regulatory agency) requirements. Also, by performing the human tissue-based tertiary study prior to commencement of human clinical studies, the potential for cross-species extrapolation errors based on animal study results has also been significantly reduced.

Prior to purchasing our in vitro human tissue equivalents, you can purchase purified total RNA (control and treated) and/or Protein Lysate (control) from each of our human tissue equivalents to confirm the expression level of specific gene(s) or the presence/absence of specific protein(s).

Q8: What do these in vitro human tissue equivalents cost?

MatTek’s in vitro human tissue equivalents are very reasonably priced, especially considering the costly materials used to produce these tissues, and the amount of time (2-3 weeks) and labor (numerous technician “touches” per lot) required to grow a given tissue. The cost of these tissues is considerably less than the cost of the end user trying to produce these tissue equivalents reproducibly from initial harvesting of seed cells. Printed price lists are available upon request from MatTek—to receive one, just complete our online Information Request Form.

Typically, because of the nature of the production process, MatTek does not provide free samples of our in vitro human tissue equivalents. MatTek will provide proof of concept to qualified potential Customers. Please contact MatTek for details.

Q13: Do you ship in vitro tissues outside the USA? Are there any special regulations or paperwork that needs to be filled out?

Yes, MatTek has many international Customers, particularly in the European Union. Each country has a unique set of shipping requirements. Please contact MatTek for more information about shipping requirements for a particular country.

Q14: We have products that need to be tested, but don't have the on-site expertise to perform these tests? Will MatTek perform the tests for us?

MatTek can do a proof of concept for your organization–please contact MatTek for details. Once we have a conversation about your intended application, we can also send you a list of MatTek-approved Contract Testing Labs that can perform assays using MatTek in vitro human tissue equivalents.

Q15: Are there detailed protocols that I can follow to use MatTek in vitro human tissue equivalents?

Yes, MatTek has detailed protocols for all of the major applications for each of our in vitro human tissue equivalents. Please contact MatTek with your specific application and we will provide the appropriate protocol for your review.

Q16: What is the significance of each of the following terms as they apply to MatTek's in vitro human tissue equivalents?

Normal Cells: Non-transformed cells. MatTek does not use immortalized cell lines. By definition, an immortalized cell line has been transformed, and therefore possesses characteristics that are not desirable in a true in vitro human tissue equivalent.Human Cell-Derived: All MatTek tissue equivalents are derived from human cells. This procedure eliminates the cross-species extrapolation concerns that accompany all work done using non-human cells.Organotypic: MatTek’s production process produces differentiated, multi-layer, 3-D human tissue equivalents that closely resemble those found in vivo. MatTek advances in tissue engineering have made it feasible to use in vitro human tissue equivalents to explore many of the scientific questions that could only be pursued in vivo previously.

Q17: What is the single most important feature of MatTek in vitro human tissue equivalents?

Reproducibility. No other feature is more important for the successful use of in vitro human tissue equivalents than reproducibility. Why? Without proven reproducibly, an organization can not use this technology in applications that are designed to run over an extended period of time. For example, if your organization plans to use in vitro human tissue equivalents to build a database of dermal irritation information about its product formulations, that database will remain useful over many years only if it is based on data from assays that use highly reproducible in vitro human tissue equivalents.

For this reason, MatTek has spent many years developing the most reproducible in vitro human tissue equivalents available. For example, MatTek has over 15 YEARS of reproducibility data for the EpiDerm human skin equivalent.

Reproducibility is so important to the successful use of in vitro human tissue equivalents that MatTek GUARANTEES the reproducibility of ALL of its in vitro tissues.

Q18: What is the best way to get specific information about my in vitro application?

Complete the MatTek Information Request Form including your detailed application information. We will review your application information and recommend the appropriate in vitro human tissue equivalent and protocol for that application. The more detailed you are in the description of how you intend to use our products in your work, the more specific we can be in our to response to your inquiry.

Q19: What are all of these TR's referenced on your Web site?

“TR” indicates a MatTek Technical Reference. Technical References are technical posters/papers that scientific staff members at MatTek and/or at organizations using MatTek in vitro products have presented at major technical conferences and meetings.

There is a searchable database of TR citations/summaries on the MatTek Web site. There are over 700 TR’s (and growing!) listed in the database. Click here to go to the MatTek TR page.

Q20: What is the quickest method to see if MatTek has information related to a specific Application, Keyword, Individual, or Organization?

As mentioned above, there is a searchable database of Technical Reference (TR) citations/summaries on the MatTek Web site. There are over 700 TR’s listed in the database. Click on this link to go to the MatTek TR page.

Q21: What are ICCVAM and ECVAM and why do I see them referenced on the MatTek Web site?

ICCVAM (USA) and ECVAM (Europe) are the organizations most directly responsible for the effort to introduce and validate non-animal alternative toxicological test methods in order to eliminate/reduce the number of animals used in these studies.

From the ICCVAM Web site: “The Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) was established in 1997 by the Director of the National Institute of Environmental Health Sciences (NIEHS) to implement NIEHS directives in Public Law (P.L.) 103-43. This law directed NIEHS to develop and validate new test methods, and to establish criteria and processes for the validation and regulatory acceptance of toxicological testing methods. P.L. 106-545, the ICCVAM Authorization Act of 2000, established ICCVAM as a permanent committee. The Committee is composed of representatives from 15 Federal regulatory and research agencies; these agencies generate, use, or provide information from toxicity test methods for risk assessment purposes. The Committee coordinates cross-agency issues relating to development, validation, acceptance, and national/international harmonization of toxicological test methods.”

From the ECVAM Web site: “ECVAM (European Centre for the Validation of Alternative Methods) was created by a Communication from the Commission to the Council and the Parliament in October 1991, pointing to a requirement in Directive 86/609/EEC on the protection of animals used for experimental and other scientific purposes, which requires that the Commission and the Member States should actively support the development, validation and acceptance of methods which could reduce, refine or replace the use of laboratory animals. ECVAM was established in 1992 as a unit of the Environment Institute, part of the Joint Research Centre, and was transferred to, at that time, the newly formed Institute for Health and Consumer Protection in Ispra, Italy in 1998.”

Q22: What is a 'validated' in vitro method? Do MatTek in vitro human tissue equivalents have validated methods?

From: The Principles and Procedures of Validation, Chapter 2 (ATLA 30, Supplement 1, 13.19, 2002): “The validation of an alternative method is the process by which the relevance and reliability of the method are established for a particular purpose. In the context of a replacement test method, relevance refers to the scientific basis of the test system, and to the predictive capacity of an associated prediction model (PM), whereas reliability refers to the reproducibility of test results, both within and between laboratories, and over time.”

The entire validation process for an alternative method can take 8-10 years. The scientific portion of the process (formal pre-validation and validation studies) usually takes 4-6 years.

MatTek has TWO alternative methods formally validated – EpiDerm for skin irritation testing and for dermal corrosion testing. Several others have completed major portions of the process – EpiOcular for ocular irritation (an alternative to the Draize Rabbit Eye Test) that is being sponsored by Colgate-Palmolive, and EpiDerm for percutaneous absorption testing.

Q23: What is the MTT Assay and what is it used for?

The MTT Assay is a very accurate end point (method) used to measure the cell viability of in vitro human tissue equivalents. The assay is a colorimetric assay that measures the reduction of a tetrazolium component (MTT) into an insoluble formazan product by the mitochondria of viable cells. After incubation of the cells with the MTT reagent for several hours, a solution is added to lyse the cells and solubilize the colored crystals. Samples are read using an ELISA plate reader at a wavelength of 570 nm. The amount of color produced is directly proportional to the number of viable cells.

Q24: Can any MatTek in vitro human tissue equivalents be used in Transdermal Drug Delivery studies?

Yes, MatTek’s EpiDerm in vitro human tissue equivalent has been used successfully in several such studies. Click on the following link to review information related to this topic: Transdermal Drug Delivery

Q25 Can any MatTek in vitro human tissue equivalents be used in Inhaled / Nasal Drug Delivery studies?

Yes, MatTek’s EpiAirway tracheal/bronchial in vitro human tissue equivalent has been used successfully in several intranasal and inhaled drug delivery studies. Click on the following links to review information related to Drug Delivery.

Q26: Where is the 'Glass Bottom Dish' Product Line information on the MatTek Web site?

MatTek’s Glass Bottom Culture Dishes are standard size 35 and 50 mm disposable plastic petri dishes with glass coverslip bottoms, providing researchers with superior high resolution microscopic images of their in vitro cultures. These dishes are routinely used in confocal, polarized light, and fluorescence imaging techniques. These dishes are also ideal for live cell microscopy.

Glass Bottom Dishes with gridded glass coverslips are also available. Part numbers include P35G-1.5-14-CGRD, P50G-1.5-14-FGRD.

MatTek Cell Culture Coverslip Kits

MatTek Cell Culture Coverslip Kits come ready to use – coverslips placed in 35 mm petri dishes and pre-sterilized. Standard glass coverslip kits are available in the CSGK/F, CSGK/C and CSGK/N Coverslip Kit configurations. These kits, the CSGK/F in particular, are widely used in amniocentesis testing.